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Pluronics-Formulated Farnesol Promotes Efficient Killing and Demonstrates Novel Interactions with Streptococcus mutans Biofilms.

Mogen AB, Chen F, Ahn SJ, Burne RA, Wang D, Rice KC - PLoS ONE (2015)

Bottom Line: In each tested S. mutans strain, biomass was significantly decreased (SNK test, p < 0.05) in the P85F and F biofilms relative to untreated biofilms.Parallel CFU/ml determinations revealed that biofilm growth in the presence of P85F resulted in a 3-log reduction in viability, whereas F decreased viability by less than 1-log.Collectively, these results suggest that Pluronics-formulated farnesol induces alterations in biofilm architecture, presumably via interaction with the sucrose-dependent biofilm matrix, and may be a viable treatment option in the prevention and treatment of pathogenic plaque biofilms.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, Florida, 32611, United States of America.

ABSTRACT
Streptococcus mutans is the primary causative agent of dental caries, one of the most prevalent diseases in the United States. Previously published studies have shown that Pluronic-based tooth-binding micelles carrying hydrophobic antimicrobials are extremely effective at inhibiting S. mutans biofilm growth on hydroxyapatite (HA). Interestingly, these studies also demonstrated that non-binding micelles (NBM) carrying antimicrobial also had an inhibitory effect, leading to the hypothesis that the Pluronic micelles themselves may interact with the biofilm. To explore this potential interaction, three different S. mutans strains were each grown as biofilm in tissue culture plates, either untreated or supplemented with NBM alone (P85), NBM containing farnesol (P85F), or farnesol alone (F). In each tested S. mutans strain, biomass was significantly decreased (SNK test, p < 0.05) in the P85F and F biofilms relative to untreated biofilms. Furthermore, the P85F biofilms formed large towers containing dead cells that were not observed in the other treatment conditions. Tower formation appeared to be specific to formulated farnesol, as this phenomenon was not observed in S. mutans biofilms grown with NBM containing triclosan. Parallel CFU/ml determinations revealed that biofilm growth in the presence of P85F resulted in a 3-log reduction in viability, whereas F decreased viability by less than 1-log. Wild-type biofilms grown in the absence of sucrose or gtfBC mutant biofilms grown in the presence of sucrose did not form towers. However, increased cell killing with P85F was still observed, suggesting that cell killing is independent of tower formation. Finally, repeated treatment of pre-formed biofilms with P85F was able to elicit a 2-log reduction in viability, whereas parallel treatment with F alone only reduced viability by 0.5-log. Collectively, these results suggest that Pluronics-formulated farnesol induces alterations in biofilm architecture, presumably via interaction with the sucrose-dependent biofilm matrix, and may be a viable treatment option in the prevention and treatment of pathogenic plaque biofilms.

No MeSH data available.


Related in: MedlinePlus

Quantification of P85-formulated farnesol effects on sucrose-independent S. mutans biofilms.Biofilms described in Fig 6 (12 random fields of view acquired from 2 independent experiments) were analyzed with COMSTAT software [34] to quantify CLSM z-stack images for biomass (A) and roughness co-efficient (B). The average CFU/ml (n = 3 biological replicates, representative of two independent experiments) of biofilms grown in each condition was also determined in a parallel experiment (C). Error bars = SEM. *Indicates statistical significance compared to untreated control •Denotes statistically-significant difference compared to farnesol-treated condition (p < 0.05; SNK test).
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pone.0133886.g007: Quantification of P85-formulated farnesol effects on sucrose-independent S. mutans biofilms.Biofilms described in Fig 6 (12 random fields of view acquired from 2 independent experiments) were analyzed with COMSTAT software [34] to quantify CLSM z-stack images for biomass (A) and roughness co-efficient (B). The average CFU/ml (n = 3 biological replicates, representative of two independent experiments) of biofilms grown in each condition was also determined in a parallel experiment (C). Error bars = SEM. *Indicates statistical significance compared to untreated control •Denotes statistically-significant difference compared to farnesol-treated condition (p < 0.05; SNK test).

Mentions: S. mutans produces an EPS biofilm matrix primarily composed of a glucan polymer when grown in the presence of sucrose, and production of this sucrose-dependent matrix is dependent on GTF enzymes [36–38]. To determine if formation of towers and/or increased cell death of P85F treated S. mutans biofilms was dependent on production of the sucrose-dependent biofilm matrix, UA159 biofilms were grown in media lacking sucrose in the presence of NBM, P85F, and F. Although formation of “death towers” was not observed in these P85F sucrose-independent biofilms (Fig 6B), they displayed decreased biomass (Fig 7A) and increased roughness (Fig 7B) relative to untreated biofilms (Fig 6D), biofilms treated with P85 (Fig 6A) or farnesol alone (Fig 6C). Additionally, S. mutans cells in both the P85F and F alone biofilms (Fig 6B and 6C) each displayed the characteristic increased cell chain length and patchy biofilm morphology that was observed in F alone biofilms grown in sucrose-containing media (Fig 1C).


Pluronics-Formulated Farnesol Promotes Efficient Killing and Demonstrates Novel Interactions with Streptococcus mutans Biofilms.

Mogen AB, Chen F, Ahn SJ, Burne RA, Wang D, Rice KC - PLoS ONE (2015)

Quantification of P85-formulated farnesol effects on sucrose-independent S. mutans biofilms.Biofilms described in Fig 6 (12 random fields of view acquired from 2 independent experiments) were analyzed with COMSTAT software [34] to quantify CLSM z-stack images for biomass (A) and roughness co-efficient (B). The average CFU/ml (n = 3 biological replicates, representative of two independent experiments) of biofilms grown in each condition was also determined in a parallel experiment (C). Error bars = SEM. *Indicates statistical significance compared to untreated control •Denotes statistically-significant difference compared to farnesol-treated condition (p < 0.05; SNK test).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4519314&req=5

pone.0133886.g007: Quantification of P85-formulated farnesol effects on sucrose-independent S. mutans biofilms.Biofilms described in Fig 6 (12 random fields of view acquired from 2 independent experiments) were analyzed with COMSTAT software [34] to quantify CLSM z-stack images for biomass (A) and roughness co-efficient (B). The average CFU/ml (n = 3 biological replicates, representative of two independent experiments) of biofilms grown in each condition was also determined in a parallel experiment (C). Error bars = SEM. *Indicates statistical significance compared to untreated control •Denotes statistically-significant difference compared to farnesol-treated condition (p < 0.05; SNK test).
Mentions: S. mutans produces an EPS biofilm matrix primarily composed of a glucan polymer when grown in the presence of sucrose, and production of this sucrose-dependent matrix is dependent on GTF enzymes [36–38]. To determine if formation of towers and/or increased cell death of P85F treated S. mutans biofilms was dependent on production of the sucrose-dependent biofilm matrix, UA159 biofilms were grown in media lacking sucrose in the presence of NBM, P85F, and F. Although formation of “death towers” was not observed in these P85F sucrose-independent biofilms (Fig 6B), they displayed decreased biomass (Fig 7A) and increased roughness (Fig 7B) relative to untreated biofilms (Fig 6D), biofilms treated with P85 (Fig 6A) or farnesol alone (Fig 6C). Additionally, S. mutans cells in both the P85F and F alone biofilms (Fig 6B and 6C) each displayed the characteristic increased cell chain length and patchy biofilm morphology that was observed in F alone biofilms grown in sucrose-containing media (Fig 1C).

Bottom Line: In each tested S. mutans strain, biomass was significantly decreased (SNK test, p < 0.05) in the P85F and F biofilms relative to untreated biofilms.Parallel CFU/ml determinations revealed that biofilm growth in the presence of P85F resulted in a 3-log reduction in viability, whereas F decreased viability by less than 1-log.Collectively, these results suggest that Pluronics-formulated farnesol induces alterations in biofilm architecture, presumably via interaction with the sucrose-dependent biofilm matrix, and may be a viable treatment option in the prevention and treatment of pathogenic plaque biofilms.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, Florida, 32611, United States of America.

ABSTRACT
Streptococcus mutans is the primary causative agent of dental caries, one of the most prevalent diseases in the United States. Previously published studies have shown that Pluronic-based tooth-binding micelles carrying hydrophobic antimicrobials are extremely effective at inhibiting S. mutans biofilm growth on hydroxyapatite (HA). Interestingly, these studies also demonstrated that non-binding micelles (NBM) carrying antimicrobial also had an inhibitory effect, leading to the hypothesis that the Pluronic micelles themselves may interact with the biofilm. To explore this potential interaction, three different S. mutans strains were each grown as biofilm in tissue culture plates, either untreated or supplemented with NBM alone (P85), NBM containing farnesol (P85F), or farnesol alone (F). In each tested S. mutans strain, biomass was significantly decreased (SNK test, p < 0.05) in the P85F and F biofilms relative to untreated biofilms. Furthermore, the P85F biofilms formed large towers containing dead cells that were not observed in the other treatment conditions. Tower formation appeared to be specific to formulated farnesol, as this phenomenon was not observed in S. mutans biofilms grown with NBM containing triclosan. Parallel CFU/ml determinations revealed that biofilm growth in the presence of P85F resulted in a 3-log reduction in viability, whereas F decreased viability by less than 1-log. Wild-type biofilms grown in the absence of sucrose or gtfBC mutant biofilms grown in the presence of sucrose did not form towers. However, increased cell killing with P85F was still observed, suggesting that cell killing is independent of tower formation. Finally, repeated treatment of pre-formed biofilms with P85F was able to elicit a 2-log reduction in viability, whereas parallel treatment with F alone only reduced viability by 0.5-log. Collectively, these results suggest that Pluronics-formulated farnesol induces alterations in biofilm architecture, presumably via interaction with the sucrose-dependent biofilm matrix, and may be a viable treatment option in the prevention and treatment of pathogenic plaque biofilms.

No MeSH data available.


Related in: MedlinePlus